College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, China.
College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, China; Key Laboratory of Animal Diseases and Human Health of Sichuan Province, Chengdu, Sichuan, China.
Vet Microbiol. 2020 Dec;251:108886. doi: 10.1016/j.vetmic.2020.108886. Epub 2020 Oct 17.
Messenger RNA-based vaccines represent new tools with prophylactic and therapeutic potential characterized by high flexibility of application for infectious diseases. Pseudorabies virus (PRV) is one of the major viruses affecting the pig industry. PRV has serious effects in piglets, sows, and growing-fattening pigs and can lead to huge economic losses. In this study, an envelope glycoprotein D (gD) gene-based specific mRNA vaccine was generated, and a mouse model was used to investigate the protective efficacy of the vaccine. The gD mRNA vaccine and the recombinant plasmid pVAX-gD were transfected into BHK21 cells, and the antigenicity of the expressed proteins was detected by Western blot analysis. Groups of mice were vaccinated with the gD mRNA vaccine, pVAX-gD, and PBS. T cell immune responses were measured by flow cytometry or ELISA and serum neutralization tests every two weeks. The challenge with the PRV-XJ strain was performed eight weeks after the primary immunization, and the response was monitored for 15 days. The levels of specific and neutralizing antibodies in the gD mRNA vaccine group were significantly increased in 8 weeks compared to those in the control group, and cytokine levels, including that of IFN-γ/IL-2, were considerably higher than those in the control animal. Additionally, the proportion of CD4/CD8 cells in peripheral lymphocytes was remarkably increased. Our data demonstrate that mRNA is a promising and effective tool for the development of vaccines. The PRV-gD-based mRNA vaccine can elicit an efficient neutralizing antibody response and induce effective protection in mice in defense against PRV infection.
基于信使 RNA 的疫苗代表了具有预防和治疗潜力的新工具，其应用具有高度的灵活性，适用于传染病。伪狂犬病病毒（PRV）是影响养猪业的主要病毒之一。PRV 对仔猪、母猪和育肥猪有严重影响，可导致巨大的经济损失。在本研究中，生成了一种基于包膜糖蛋白 D（gD）基因的特异性 mRNA 疫苗，并使用小鼠模型来研究疫苗的保护效力。将 gD mRNA 疫苗和重组质粒 pVAX-gD 转染到 BHK21 细胞中，通过 Western blot 分析检测表达蛋白的抗原性。将 gD mRNA 疫苗、pVAX-gD 和 PBS 分别接种到小鼠组中。每两周通过流式细胞术或 ELISA 检测 T 细胞免疫反应，并进行血清中和试验。在初次免疫后 8 周进行 PRV-XJ 株的攻毒，监测反应 15 天。与对照组相比，gD mRNA 疫苗组在 8 周时特异性和中和抗体水平显著升高，IFN-γ/IL-2 等细胞因子水平明显升高。此外，外周淋巴细胞中 CD4/CD8 细胞的比例显著增加。我们的数据表明，mRNA 是疫苗开发的一种有前途且有效的工具。基于 PRV-gD 的 mRNA 疫苗能够在小鼠中诱导有效的中和抗体反应，并在针对 PRV 感染的防御中产生有效的保护作用。
